转化生长因子β1诱导肾间质成纤维细胞表型转化及其对结缔组织生长因子基因表达的影响  

The experimental study of the effects of renal interstitial fibroblast cells activation induced by TGF-β1 on ctgf gene expression

作　　者：周瑾[1] 陈香美[1] 刘述文[1] 吕杨[1] 傅博[1] 

机构地区：[1]中国人民解放军总医院肾病中心暨国家重点实验室,北京100853

出　　处：《临床肾脏病杂志》2014年第5期299-303,共5页Journal Of Clinical Nephrology

基　　金：国家自然科学基金重点资助课题（No.30130220）及面上项目（No.30873345）资助

摘　　要：目的 研究转化生长因子β1（transforming growth factor beta-1,TGF-β1）诱导大鼠正常肾间质成纤维细胞（NRK-49F）表型转化过程中结缔组织生长因子（connective tissue growth factor,CTGF）基因表达的变化.方法 以不同浓度的TGF-β1（0、0.5、1、2、5、10 ng/ml）刺激NRK-49F细胞,分别应用MTT比色法、Western Blot、Northern Blot方法,检测TGF-β1刺激后肾间质成纤维细胞的增殖、Ⅰ型、Ⅲ型前胶原mRAN的表达、细胞表型标志物α-SMA mRNA及蛋白质表达、CTGFmRNA的表达变化.结果 1 ng/ml浓度以上TGF-β1能显著促进NRK-49F细胞增殖,上调Ⅰ、Ⅲ型前胶原mRNA的表达水平,诱导肌成纤维细胞表型标志α-SMA mRNA及蛋白质的表达,显著上调CTGF mRNA的表达水平（P＜0.01或P＜0.05）;以上效应均呈浓度依赖性.同时CTGF mRNA表达水平的增高与细胞表型转化的程度相一致.结论 TGF β1能诱导肾间质成纤维细胞发生表型转化,并促进了细胞增殖及细胞外基质的合成;该效应与TGF β1显著上调CTGF的基因表达一致.Objective To investigate the effects of renal interstitial fibroblast cells activation induced by transforming growth factor beta-1 （TGF-β1）on cornective tissue growth factor（CTGF） gene expression.Methods Normal rat renal interstitial fibroblast cells （NRK 49F） were induced by hTGFβ1 at the different concentrations of 0.5ng/ml,1ng/ml,2ng/ml,5ng/ml,10ng/ml,respectively,in contrast with phosphatic buffered saline,MTT assay was used to measure the proliferation of NRK49F,Western Blot and Northern Blotting were used to measure the expression of alpha-smooth muscle actin （α-SMA）,and the mRNA levels of type Ⅰ and Ⅲ procollagen and CTGF were also detected.Results TGF-β1 at the concentration as low as 2-10 ng/ml could promote NRF-49F proliferation,increased the gene expression of Ⅰ and Ⅲ procollagen,boosted α-SMA protein and gene expression which represented myofobroblast appearance in concentration-dependant form,at the same time enhanced the gene expression of CTGF,which were compatible with interstitial fibroblast cells phenotype differentiation.Conclusions TGF-β1 could not only induce renal interstitial fibroblast activation,but also promote its proliferation and sythesise of extracellular matrix（ECM）,whose effects were related with the CTGF gene expression up-regulation.

关 键 词：转化生长因子Β1 肾间质成纤维细胞 表型转化 细胞外基质 结缔组织生长因子 

分 类 号：R692[医药卫生—泌尿科学]